ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2014, Vol. 45 ›› Issue (8): 1324-1329.doi: 10.11843/j.issn.0366-6964.2014.08.017

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Construction of Bacterial Artificial Chromosome (BAC) Library of Ovine Babesia motasi

ZHANG Hao-hao1,3,CHEN Long-xin2,3,CAO Yang3,CHEN Ying3,GUAN Gui-quan1*,MA Run-lin2,3*   

  1. (1.State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;2.Zhengzhou Normal University,Zhengzhou 450044,China;3.Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Beijing 100101,China)
  • Received:2014-01-25 Online:2014-08-23 Published:2014-08-23

Abstract:

 To enrich the genomic information,the merozoites of ovine Babesia motasi Lintan were embedded in low-melting-point (LMP) agarose and subsequently digested with proteinase K to get their high-quality megabase size genomic DNA.Then the large size DNA fragments isolated by pulse-field gel electrophoresis (PFGE) were ligated to CopyControl pCC1BAC Cloning-ready Vector.The present results showed that the bacterial artificial chromosome (BAC) library containing 34 560 clones was successfully constructed by transformation of the ligated DNA into EPI300TM Escherichia coli.In B.motasi Lintan BAC library,the average length of the insert DNA fragment was 70 kb and covers ×137 B.motasi Lintan genome.The BAC library in which the empty clone was less than 3% and the clones were stable should contribute to fine physical mapping,genome sequencing,and evolution and virulence genes derived from comparative genomics of Babesia species and so on.

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